Photoaffinity labeling of D1 and D5 dopamine receptors.
Although human D1 and D5 dopamine receptors are encoded by distinct genes and share only 50% sequence homology at the amino acid level, their pharmacological properties are identical. Using a selective D1 receptor photoaffinity radioligand, (+/-)-7-[125I]iodo-8-hydroxy-3-methyl-1-(4-azidophenyl)-2,3,4,5-tetrahyd ro-1H-3-benzazepine ([125I]MAB), we have further probed the molecular properties of these receptors in transfected GH4C1 rat pituitary cells. Under reversible, non-covalent binding conditions, [125I]MAB bound to both the D1 and the D5 receptors with identical affinities, dopaminergic selectivity and stereospecificity. Upon photoactivation of the bound [125I]MAB, the label was incorporated into a approximately 64,000 mol. wt protein corresponding to the D1 dopamine receptor. However, there was no specific photoincorporation of the ligand observed in D5 receptors. The lack of [125I]MAB photolabeling of D5 receptors was independent of the cell line chosen, since similar results were obtained using other transfected cells. The data suggest that although both D1 and D5 receptors share structurally similar binding sites, the protein domains around the sites are different. Thus, although there are currently no specific compounds which bind preferentially to D1 or D5 receptors, these receptors can be distinguished from one another by the inability of [125I]MAB to photolabel D5, but not D1, receptors. Such selective targeting of a specific receptor may be useful in understanding the functional importance and/or interaction between closely related members of the same receptor family when co-expressed in the same cell.
Sidhu
A
A
Department of Pediatrics, Georgetown University Medical Center, Georgetown University, Washington, DC 20007, USA.
Banta
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M
Uh
M
M
Shin
Y
Y
D000818:Animals; D001386:Azides; D001552:Benzazepines; D046508:Culture Techniques; D006801:Humans; D007457:Iodine Radioisotopes; D008566:Membranes / Q000378:metabolism; D008970:Molecular Weight; D019920:Photoaffinity Labels; D010902:Pituitary Gland / Q000166:cytology / Q000378:metabolism; D011869:Radioligand Assay; D051381:Rats; D017447:Receptors, Dopamine D1 / Q000378:metabolism*; D050641:Receptors, Dopamine D5; D014162:Transfection
D001386:Azides; D001552:Benzazepines; C493605:DRD5 protein, human; C493607:Drd5 protein, rat; D007457:Iodine Radioisotopes; D019920:Photoaffinity Labels; D017447:Receptors, Dopamine D1; C058320:7-iodo-8-hydroxy-3-methyl-1-(4-azidophenyl)-2,3,4,5-tetrahydro-1H-3-benzazepine; D050641:Receptors, Dopamine D5